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DNA ligation is the procedure of binding a DNA fragment into a vector backbone with a covalent bond (a phosphodiester link) and is usually performed employing the T4 DNA ligase enzyme. The procedure requires the molecules of DNA to have compatible single stranded overhangs (sticky ends). Therefore, both the fragment and the vector should be first digested with the same restriction enzymes. If only one restriction enzyme is used in the procedure, make sure to treat the linearized vector backbone with phosphatases before proceeding. The use of phosphatase-treated vectors in the ligation step...